Rescue of very virulent and mosaic infectious bursal disease virus from cloned cDNA: VP2 is not the sole determinant of the very virulent phenotype

被引:121
作者
Boot, HJ [1 ]
Ter Huurne, AAHM [1 ]
Hoekman, AJW [1 ]
Peeters, BPH [1 ]
Gielkens, ALJ [1 ]
机构
[1] Inst Anim Sci & Hlth, Dept Avian Virol, Lelystad, Netherlands
关键词
D O I
10.1128/JVI.74.15.6701-6711.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Many recent outbreaks of infectious bursal disease in commercial chicken flocks worldwide are due to the spread of very virulent strains of infectious bursal disease virus (vvIBDV). The molecular determinants for the enhanced virulence of vvIBDV compared to classical IBDV are unknown. The lack of a reverse genetics system to rescue vvIBDV from its cloned cDNA hampers the identification and study of these determinants. In this report we describe, fur the first time, the rescue of vvIBDV from its cloned cDNA, Two plasmids containing a T7 promoter and either the full-length A- or B-segment cDNA of vvIBDV (D6948) were cotransfected into QM5 cells expressing T7 polymerase, The presence of vvIBDV could be detected after passage of the transfection supernatant in either primary bursa cells (in vitro) or embryonated eggs (in vivo), but not QM5 cells. Rescued vvIBDV (rD6948) appeared to have the same virulence as the parental isolate, D6948, Segment-reasserted IBDV, in which one of the two genomic segments originated from cDNA of classical attenuated IBDV CEF94 and the other from D6938, could also be rescued by using this system, Segment-reasserted virus containing the A segment of the classical attenuated isolate (CEF94) and the B segment of the very virulent isolate (D6948) is not released until 15 h after an in vitro infection, This indicates a slightly retarded replication, as the first release of CEF94 is already found at 10 h after infection, Next to segment reassortants, we generated and analyzed mosaic IBDVs (mIBDVs), In these mIBDVs we replaced the region of CEF94 encoding one of the viral proteins (pVP2, VP3, or VP4) by the corresponding region of D6948, Analysis of these mIBDV isolates showed that tropism for non-B-lymphoid cells was exclusively determined by the viral capsid protein VP2, However, the very virulent phenotype was not solely determined by this protein, since mosaic virus containing VP2 of vvIBDV induced neither morbidity nor mortality in young chickens.
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收藏
页码:6701 / 6711
页数:11
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